In this year of our study, we will concentrate on the determination of the amino acid sequence of the glycopeptide repeating segments of bovine submaxillary glycoprotein. These segments, which are approximately 28 amino acids in length, constitute a family of glycopeptides whose members differ from one another in the positions of several amino acids. We have referred to these differences as homologous substitutions. The primary concern for the first part of the year will be to effect a separation of the members of the homologous glycopeptide family. We will initially use classical chromatographic media, such as Dowex-50, Dowex-1, phosphocellulose, and ion exchange cellulose for separation. Where possible HPLC methodology will be used to obtain maximum resolution. An LKB Ultragrad will be used for non-linear gradient elution. Columns of spherical hydroxylapatite and ODS reverse phase media will also be tried. The separated segments will then be sequenced by the Edman procedure, if necessary, using a micro-modification we have developed based on fluorescence. Peptides obtained through beta-elimination and hydrolysis will be used to provide overlapping sequences. BIBLIOGRAPHIC REFERENCES: A. Wu and W. Pigman. Preparation and Characterization of Armadillo Submandibular Glycoproteins. Biochem. J., 166, 37-47 (1977). F. Downs and W. Pigman. Determination of O-Glycosidic Linkages to L-Serine and L-Threonine Residues of Glycoproteins. Methods in Carbohydrate Chemistry, Vol. VII, R.L. Whistler and J.N. BeMiller, editors, pp. 200-204, Academic Press, New York (1976).